Energy-saving pretreatments affect pelagic Sargassum composition and DNA metabarcoding reveals the microbial community involved in methane yield.

Sargassum spp. flood the Caribbean coastline, causing damage to the local economy and environment. Anaerobic digestion (AD) has been proposed as an attractive option for turning macroalgae into valuable resources. Sargassum spp. has a complex composition that affects the microbial composition involved in AD which generates a low methane yield. This study aimed to improve the methane yield of pelagic Sargassum, using different energy-saving pretreatments and identifying the microbial community associated with methane production. We applied different energy-saving pretreatments to algal biomass and assessed the methane yield using a biomethane potential (BMP) test. The microbial communities involved in the AD of the best- and worst-performing methanogenic systems were analyzed by high-throughput sequencing. The results showed that pretreatment modified the content of inorganic compounds, fibers, and the C:N ratio, which had a strong positive correlation with BMP. The water washing pretreatment resulted in the best methane yield, with an increase of 38%. DNA metabarcoding analysis revealed that the bacterial genera Marinilabiliaceae_uncultured, DMER64, Treponema, and Hydrogenispora, as well as the archaea genera Methanosarcina, RumEn_M2, Bathyarchaeia, and Methanomassiliicocus, dominated the microbial community with a high methane yield. This study is the first to demonstrate the microbial community structure involved in the AD of Sargassum spp. The pretreatments presented in this study can help overcome the limitations associated with methane yield.

Synergy of the Tropical Earthworm Pontoscolex corethrurus and Oil Palm Bagasse in the Removal of Heavy Crude Oil.

The tropical endogeic earthworm Pontoscolex corethrurus, a non-standard species used in ecotoxicity, has been found in crude oil-contaminated habitats. We estimated the removal of total hydrocarbons from heavy crude "Maya" oil on an artificially contaminated soil with a median lethal concentration of P. corethrurus and an addition of oil palm bagasse. P. corethrurus had a high survival rate, and the addition of oil palm bagasse led to a greater growth and an increase in abundance of bacteria and fungi. The activity of P. corethrurus and the nutritional quality of oil palm bagasse had a significant impact on the removal of a larger amount of petroleum hydrocarbons from contaminated soil. We concluded that the endogeic earthworm P. corethrurus and oil palm bagasse acted synergistically to achieve a more effective removal of total petroleum hydrocarbons from soil. These results show the potential for using P. corethrurus to remove, either directly or indirectly, crude oil from soil.

Methane Production of Sargassum spp. Biomass from the Mexican Caribbean: Solid-Liquid Separation and Component Distribution.

In the last decade, Sargassum spp. seaweed species have caused massive flooding on the Caribbean Sea coasts. These seaweed species have a high content of recalcitrant compounds, such as insoluble fibers and polyphenols, which generate low methane yields in anaerobic digestion (AD). This study investigated the effect of solid-liquid separation of Sargassum biomass on biodegradability and methane yield. A biochemical methane potential (BMP) test was conducted with both fractions and raw biomass (RB). A mass balance was developed to assess the distribution of the components. The obtained liquid fraction (LF) showed high biodegradability and a high methane production rate, and it generated a methane yield of 159.7 ± 7.1 N L kg VS-1, a value that corresponds to approximately twice that achieved with RB and the solid fraction (SF). The component distribution analysis showed that about 90% of total solids (TS), volatile solids (VS), ash, carbon, and cellulose were retained in the SF. In conclusion, the LF had high biodegradability and methane yield. This suggests the potential for LFs of Sargassum biomass to be treated in large-scale high-load reactors; however, studies applied to SFs are needed because they retain a large amount of organic matter with low biodegradability.

Physiological stabilization, community characterization, and nitrogen degradation dynamics in an anammox consortium from estuarine sediments.

Anammox is a cost-effective and sustainable process for nitrogen removal; however, the production of a physiologically stable inoculum is a critical point in the start-up process. In this work, estuarine sediments were used as incubation seeds to obtain cultures with stable anammox activity. Assays were performed in batch cultures fed with stoichiometric amounts of ammonium and nitrite, analyzing physiological response variables and the microbial community. Estuarine sediments showed a stable anammox process after 90 days, consuming ammonium and nitrite simultaneously with concomitant generation of N2 and nitrate in stoichiometric amounts. In kinetic assays, substrates were fully consumed after 210 hr, exhibiting N2 and nitrate yields of 0.85 and 0.10, respectively. The microbial community analysis using PCR-DGGE indicated the presence of uncultured anammox bacteria and members of the genus Candidatus Jettenia. The results evidenced the achievement of anammox cultures, although their start-up and kinetic characteristics were less favorable than those recorded in man-made systems. PRACTITIONER POINTS: Estuarine sediments were used as incubation seeds to obtain cultures with stable anammox activity. The sediments were fed with stoichiometric amounts of ammonium and nitrite, analyzing the physiological response variables and the microbial community. Sediments showed a stable anammox process after 90 days, converting the substrates into N2 and nitrate according to stoichiometry. Anammox cultures were achieved although their start-up and kinetic characteristics were less favorable than those recorded in man-made systems. Microbial community analysis using PCR-DGGE indicated the presence of uncultured anaerobic ammonia-oxidizing bacterium and members of genus Candidatus Jettenia.

2-Chlorophenol consumption by cometabolism in nitrifying SBR reactors.

Cometabolic consumption of 2-chlorophenol (2-CP) by a nitrifying sludge was evaluated in two SBR reactors fed with 60 mg 2-CP-C/L and different initial ammonium concentrations (100, 200, 300, 400, and 500 mg NH4+-N/L). Irrespectively to the increase in ammonium concentration and throughout the operational cycles, the sludge achieved a complete nitrification in 14 days, accounting for ammonium consumption efficiencies close to 99% and nitrate production yields between 0.93 and 0.99. The sludge was able to completely consume 2-CP within 7 days. The increase in ammonium concentration provoked an increment in the specific rates of both ammonium (qNH4+-N) and 2-CP (q2-CP-C) consumption up to 5.2 and 3.1 times, respectively. The cometabolic effect of the increase in ammonium concentration on 2-CP consumption was supported by a direct and significant relationship between the qNH4+-N and q2-CP-C (r = 0.83). Moreover, batch assays conducted with ammonium, 2-CP, allylthiourea as specific inhibitor of the ammonium monooxygenase (AMO) enzyme, and the sludge inoculated into the reactors, resulted in a decrease of 34% in q2-CP-C, evidencing the participation of the AMO in the consumption of 2-CP. When the same assays were carried out with the sludge obtained from the SBR reactors after 13 operating cycles, a higher participation of the AMO in 2-CP consumption was noticed with a decrease of 53% in q2-CP-C. According to these results, the use of nitrifying sludge and high ammonium concentrations in SBR systems can be a suitable alternative for increasing the cometabolic consumption of recalcitrant compounds like 2-CP.

Sensitivity of the Endogeic Tropical Earthworm Pontoscolex corethrurus to the Presence of Heavy Crude Oil.

Contamination of soil with petroleum is common in oil-producing areas across the tropical regions of the world. There is limited knowledge on the sensitivity of endogeic tropical earthworms to the contamination of soil with total petroleum hydrocarbons (TPH) present in crude oil. Pontoscolex corethrurus is a dominant species in tropical agroecosystems around oil-processing facilities. The sensitivity of P. corethrurus to soil artificially contaminated with "Maya" Mexican heavy crude oil was investigated through avoidance and acute ecotoxicity tests, using the following measured concentrations: 0 (reference soil), 551, 969, 4845, 9991 and 14,869 mg/kg. The avoidance test showed that P. corethrurus displayed a significant avoidance behavior to heavy crude oil at a concentration of 9991 mg/kg or higher. In contrast, acute toxicity tests indicate that the median lethal concentration (LC50) was 3067.32 mg/kg; however, growth (weight loss) was more sensitive than mortality. Our study revealed that P. corethrurus is sensitive to TPH, thus highlighting the importance of P. corethrurus for petroleum ecotoxicological tests.

Rheological behavior of physicochemical sludges during methanogenesis suppression and hydrogen production at different organic loading rates.

This study investigated the rheological behavior of raw physicochemical sludges and sludges that were digested at different organic loading rates (OLRs) (1, 5, 10 and 15 gVS L(-1) d(-1)) during methanogenesis suppression to produce hydrogen anaerobically. The Herschel-Bulkley model was used to describe the rheology of these sludges with specific properties. The results indicate that the Herschel-Bulkley model adequately described the rheology (τ0 ≠ 0) of this type of fluids (R(2) > 0.98). In addition, the raw physicochemical sludges and those that were digested at different OLRs had dilatant behaviors (n > 1), which increased with increasing OLR. These results identified the apparent viscosity, yield stress, pH and OLR conditions that allow for the production and suppression of methane, as well as the conditions that guarantee the production of hydrogen.

Dynamics of a microbial community exposed to several concentrations of 2-chlorophenol in an anaerobic sequencing batch reactor.

The aim of this study was to contribute to the knowledge on the dynamic of the microbial community involved in anaerobic degradation of different concentrations of 2-chlorophenol (2CP, from 28 to 196 mg 2CP-C/L) and a mixture of 2CP and phenol (from 28 to 196 mg phenol-C/L) and its relationship with the respiratory process in two anaerobic sequencing batch reactors (ASBR). The dynamic of the microbial community was evaluated by denaturant gradient gel electrophoresis (DGGE) and ecological indices (S and J indices). The respiratory process was evaluated by means of substrate consumption efficiency, biogas yield, and specific consumption rates as response variables. The high consumption efficiency (90%) and the constant biogas yields obtained at concentrations up to 140 mg C/L may be related with the evenness of microbial populations (J index=0.97±0.2) present in both reactors. Pseudomonas genus was present in all concentrations tested, suggesting a possible relationship with the dehalogenation observed in both reactors. The decrease in specific consumption rate and biogas yield as well as the accumulation of phenol and volatile fatty acids observed in both reactors at 196 mg 2CP-C/L might be associated with the disappearance of the bands related to Caulobacter and Bacillus. At these conditions, the disappearance of fermentative or acetogenic bacteria resulted in reduction of substrates required to carry out methanogenesis, which eventually might cause the declination in methanogenic populations present in the reactors.

Influence of organic loading rate on methane production in a CSTR from physicochemical sludge generated in a poultry slaughterhouse.

The influence of the increase of the organic loading rate (OLR) on methane production in a continuous stirred-tank reactor (CSTR) from physicochemical sludge generated in a poultry slaughterhouse was evaluated. Total solid (TS) to obtain OLR of 1, 5, 10 and 15 g VS L(-1) day(-1), with hydraulic retention times of 29, 6, 6 and 4, respectively, were conditioned. The results showed a decrease in pH levels and an increase in the theoretical volatile fatty acids (VFA). While the yield of methane production decreased from 0.48 to 0.10 LCH4/g VSremoved, respectively, the OLR-10 managed on average 38% removal of volatile solids (VS) and a yield biogas production of 0.81 Lbiogas g(-1) VSremoved and 1.35 L day(-1). This suggests that the OLR increases in an anaerobic system from physicochemical sludge only inhibits the methanogenic metabolism, because there is still substrate consumption and biogas production.

Effect of phenol and acetate addition on 2-chlorophenol consumption by a denitrifying sludge.

Chlorophenols are widely distributed in the environment. Various strategies have been used to improve their biological elimination under anaerobic conditions; however, such information is still scarce. The aim of this study was to evaluate in batch assays the consumption of 2-chlorophenol (2-CP) by a denitrifying sludge and the influence of acetate or phenol as co-substrates in the 2-CP consumption. It was observed that phenol (69 and 92 mg phenol-C L(-1)) and acetate (60 and 108 mg acetate-C L(-1)) enhanced 2-CP consumption by the denitrifying sludge, increasing both the efficiency (up to 100%) and specific rate of 2-CP consumption. When phenol was added at 92 mg C L(-1), the specific consumption rate of 2-CP increased 2.6 times with respect to the control lacking co-substrates, whereas with acetate (108 mgC L(-1)) the increase was 9.0 times. Acetate appeared to be a better co-substrate for 2-CP consumption, obtaining a specific consumption rate of 2.48 +/- 0.14 mg 2-CP-C g(-1) VSS d(-1) at 108 mg acetate-C L(-1). The mass balance analysis indicated that the denitrifying sludge was able to simultaneously mineralize 2-CP, phenol or acetate (E2-CP, E(Phenol), and E(Acetate) close to 100% [E = consumption efficiency], Y(HCO3-) of 0.90 +/- 0.10 [Y = yield]) and reduce nitrate to nitrogen gas (E(NO3-) of 100% and Y(N2) of 0.96 +/- 0.02). It was shown that the addition of co-substrates as phenol or acetate could be a good alternative for improving the elimination of chlorophenols from wastewaters by denitrifying sludges.

2-Chlorophenol consumption and its effect on the nitrifying sludge.

The kinetic behavior of a nitrifying sludge exposed to 2-chlorophenol (2-CP) was evaluated in batch culture. The assays were performed using a stabilized nitrifying sludge. In control assays with (mg L(-1)): NH(4)(+)-N (100) and NaHCO(3)(-)-C (250), the substrates were consumed in 8h, the ammonium consumption efficiency was 99% and the NO(3)(-) yield higher than 0.9. When 5mg 2-CP-C L(-1) was added, it was transformed into an unidentified intermediate and the nitrifying efficiency decreased to 10%. Ammonium specific consumption rate diminished 95%, but the NO(3)(-) yield remained higher than 0.9. The biomass previously exposed to 2-CP was newly suspended with NH(4)(+)-N or NO(2)(-)-N in order to evaluate the ammonium and nitrite oxidizing processes. The consumption efficiencies and NO(3)(-) yields were similar to those obtained in control assays. However, the total time required for ammonium and nitrite consumption increased to 120 and 42 h, respectively. Specific consumption rates for NH(4)(+)-N and NO(2)(-)-N decreased by 95% and 83% respectively, compared to control assays. Thus, the previous contact to 2-CP had more influence on ammonium oxidizing process than the nitrite oxidizing process. These are the first evidences where a nitrifying sludge exposed to 2-CP are reported.

Acetate enhances the specific consumption rate of toluene under denitrifying conditions.

Toluene is usually present in the environment as a contaminant along with other carbon sources which may influence its removal. In this work we studied the effect of a readily consumable carbon source such as acetate on toluene mineralization under denitrifying conditions. Continuous and batch cultures with stabilized denitrifying sludge were carried out. An upflow anaerobic sludge blanket reactor (UASB) was fed with several ratios of acetate-C/toluene-C loading rates (mg C/L-day: 100/0, 75/25, 50/50, and 0/100). Batch assays with different acetate-C/toluene-C ratios (10/70, 30/50, 50/30, and 65/20 mg C/L) were also done. As the acetate loading rate decreased in the culture, the carbon and nitrate consumption efficiency decreased by 40% and 34%, respectively. HCO(3) (-) and N(2) yields also decreased by 43%. Analysis of the denitrifying community using the denaturing gradient gel electrophoresis technique indicated that there was no clear relationship between its population profile and the metabolic pattern. In batch assays, when the acetate concentration was higher than that of toluene (65 mg acetate-C/L vs 20 mg toluene-C/L), the specific consumption rate of toluene (q(T)) was two times higher than in assays with 20 mg toluene-C/L as the sole electron source (0.006 mg C/mg volatile suspended solids-day). It is proposed that acetate can act by enhancing the growth of microbial populations and as a biochemical enhancer. The results show that acetate addition can be useful to improve the consumption rate of toluene in contaminated water.